JP2015517524A - (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4 Method of using 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide - Google Patents

Abstract

A method of treating bacterial infections, for example bacterial infections caused by methicillin-resistant Staphylococcus aureus, Helicobacter pylori, Chlamydia trachomatis or Chlamydia pneumonia , Subject to (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo Disclosed is a method comprising administering -1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof. Specifically, the subject is given (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1 , 11-Dioxo-1,4,4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide crystalline monohydrochloride, crystalline monomesylate or crystalline monosulfate A method for treating a bacterial infection is disclosed. [Selection] Figure 1

Description

(Cross-reference of related applications)
This application claims the benefit of priority from US Provisional Application No. 61 / 646,754, filed May 14, 2012, the contents of which are hereby incorporated by reference in their entirety.
(Field of Invention)
The present disclosure relates to a method of treating bacterial infections or diseases associated with such infections (e.g., peptic ulcers and chlamydia), wherein the method comprises (4S, 4aS, 5aR, 12aS)- 4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11, Administering 12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof, wherein the cell infection is methicillin-resistant Staphylococcus aureus, Helicobacter pylori (Helicobacter pylori), Chlamydia trachomatis and Chlamydia pneumoniae are caused by bacteria selected from the group. In certain embodiments of the invention, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1 , 11-dioxo-1,4,4a, 5,5a, 6,11,12a-crystalline monohydrochloride, monomesylate or monosulfate of octahydro-naphthacene-2-carboxylic acid amide is administered.

  Tetracyclines are known as “broad spectrum” antibiotics and are becoming widely used for therapeutic purposes. Tetracycline has been found to be highly pharmacologically effective against rickettsia; many gram-positive and gram-negative bacteria; and pathogens that cause inguinal lymphogranulomas, inclusion body conjunctivitis, and parrot disease Yes. The first use of tetracycline antibiotics dates back to 1948. Examples of pharmaceutically active tetracycline and tetracycline analog compositions can be found in the following US patents: US Pat. Nos. 2,980,584; 2,990,331; 3,062,717; 3,165,531; 3,454,697; 3,557,280 No. 3,674,859; No. 3,957,980; No. 4,018,889; No. 4,024,272; and No. 4,126,680. Tetracycline can also be used to treat inflammatory skin diseases such as dermatitis, psoriasis, pyoderma gangrenosum, acne and rosacea.

  Methicillin-resistant Staphylococcus aureus (MRSA) is a strain of Staphylococcus aureus that is resistant to several commonly used antibiotics. In general, MRSA infections occur in hospitals and other medical facilities in people with weak immunity (nosocomial MRSA), but sometimes in otherwise healthy individuals outside the hospital Sometimes (communicable MRSA). Most MRSA infections affect the skin, but some can become severe and can spread to the bloodstream, heart or lungs, urine, or at a recent surgical site. MRSA is generally treated with antibiotics such as doxycycline, clindamycin, linezolid, or a combination of trimethoprim / sulfamethoxazole (TMP / SXT).

  Chlamydia is a sexually transmitted disease caused by the bacterium Chlamydia trachomatis. It is the most common sexually transmitted disease in the United States and causes more than 3 million cases of cervicitis and urethritis each year. Chlamydia infections in women often lead to inflammation of the cervix and can cause infertility if left untreated. Infections in men often cause inflammation of the urethra. Early treatment with Chlamydia antibiotics can prevent the development of these long-term complications.

  Chlamydia pneumoniae is a small gram-negative bacterium that often causes community-acquired respiratory infections such as pneumonia and bronchitis in adults and children.

  Peptic ulcers are diseases characterized by inflammation of the stomach and duodenal mucosa, destroying the protective mucosa of these organs, allowing stimulation by the acid produced in the stomach. A common cause of protective mucosal destruction is infection by Helicobacter pylori. Typical treatment includes the combination of antibiotics and other drugs such as proton pump inhibitors (eg omeprazole) and / or bismuth.

  Since the widespread use of tetracyclines for both major and minor illnesses resulted in resistance to these antibiotics, substituted tetracycline compounds have become bacterial infections, inflammation, neoplasms, and other diseases Developed to treat. The term “tetracycline compound” includes many compounds having a ring structure similar to tetracycline. Examples of such tetracycline compounds include: chlortetracycline, doxycycline, minocycline, oxytetracycline, demeclocycline, metacycline, sancycline, kerocardine, loritetracycline, limecycline, apiccycline; chromocycline, guamecycline, meggle Cyclin, mepyr cyclin, penimepi cyclin, piper cyclin, etamocycline, penimo cyclin. For example, substituted tetracycline compounds are disclosed in WO 2008/079339 and WO 2008/079363.

  One substituted tetracycline compound is (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11 -Dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide, which is described in U.S. Patent Application Publication Nos. 2008/0312193 and 2010/0305072. Have been described.

  In part because of the development of resistance to certain antibiotics, there is a need in the art for new tetracycline antibiotics for the treatment of diseases and disorders.

  The present invention is directed to a novel method for treating bacterial infections or diseases associated with such infections (e.g., peptic ulcer), wherein the method comprises: (4S, 4aS , 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5, Administering 5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof.

  The present invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl ] Bacteria comprising administering a 1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof Directed to a method of treating an infection, wherein the bacterium is selected from the group consisting of methicillin-resistant Staphylococcus aureus (MRSA), Helicobacter pylori, Chlamydia trachomatis, and Chlamydia pneumoniae. The invention also relates to (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy () in the manufacture of a medicament for treating bacterial infections. Use of (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof The bacterium is selected from the group consisting of methicillin-resistant Staphylococcus aureus (MRSA), Helicobacter pylori, Chlamydia trachomatis, and Chlamydia pneumoniae. In certain embodiments, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11- A crystalline salt of dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide is administered, preferably the crystalline salt is monohydrochloride, monomesylic acid It can be selected from the group consisting of salts and monosulfates.

  In a specific embodiment, the MRSA is community-associated MRSA (MRSA-CA) or hospital acquired MRSA (MRSA-HA).

  In another embodiment, the present invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl ) Amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof To a method for treating peptic ulcer. The present invention also provides (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy () in the manufacture of a medicament for treating peptic ulcer. Use of (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof About. In one embodiment, the salt is a crystalline salt and may preferably be selected from the group consisting of monohydrochloride, monomesylate, and monosulfate. In one embodiment, the method of treating peptic ulcer further comprises administering at least one additional active ingredient, such as a proton pump inhibitor and / or bismuth.

FIG. 1 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1,4,4a, 5,5a, 6,11,12a-After synthesis of crystalline monohydrochloride of octahydro-naphthacene-2-carboxylic acid amide and stored for 7 days at 40 ° C and 75% relative humidity (RH) The powder X-ray diffraction (XRPD) analysis will be shown later. FIG. 2 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1 is a differential scanning calorimetry (DSC) curve after synthesis of crystalline monohydrochloride of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide. FIG. 3 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1 is a thermogravimetric analysis (TGA) curve after synthesis of crystalline monohydrochloride of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide. FIG. 4 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- XRPD after synthesis of crystalline monomesylate of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide and after storage for 7 days at 40 ° C. and 75% RH Analysis is shown. FIG. 5 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1 is a DSC curve after synthesis of crystalline monomesylate of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylate. FIG. 6 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1,4,4a, 5,5a, 6,11,12a-TGA after synthesis of crystalline monomesylate salt of octahydro-naphthacene-2-carboxylic acid amide. FIG. 7 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- XRPD after synthesis of crystalline monosulfate of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide and after storage for 7 days at 40 ° C. and 75% RH Analysis is shown. FIG. 8 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1 is a DSC curve after synthesis of crystalline monosulfate of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide. FIG. 9 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1,4,4a, 5,5a, 6,11,12a-TGA after synthesis of crystalline monosulfate of octahydro-naphthacene-2-carboxylic acid amide. FIG. 10 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- XRPD analysis of 1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide amorphous bishydrochloride is shown. FIG. 11 shows (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1,4,4a, 5,5a, 6,11,12a—A superposition of the TGA and DSC curves of amorphous bishydrochloride of octahydro-naphthacene-2-carboxylic acid amide.

Method of the Invention A first embodiment of the invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7- [(Methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof Is directed to a method for treating a bacterial infection comprising administering a salt, wherein the bacterium is methicillin-resistant Staphylococcus aureus (MRSA), Helicobacter pylori, or Chlamydia trachomatis . In a preferred embodiment, the method provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl Administering a crystalline salt of) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide. Preferably, the crystalline salt may be selected from the group consisting of monohydrochloride, monomesylate and monosulfate. These crystalline salts are fully described in copending US application Ser. No. 13 / 471,275, the contents of which are hereby incorporated by reference in their entirety.

  In certain embodiments, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1 used , 11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof, the amount is about 10 mg to about 2000 mg, preferably Is about 25 mg to about 500 mg. In certain embodiments, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11- Dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof is at least once a month, preferably once a week, Preferably administered twice a week, most preferably daily.

  In one embodiment, the invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl ) Amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof To a method for treating methicillin-resistant Staphylococcus aureus (MRSA). In an embodiment, the invention is directed to a method of treating MRSA, wherein said MRSA is nosocomial (MRSA-HA) or community-acquired (MRSA-CA).

  In another embodiment, the invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy ( Methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof Directed to a method of treating Helicobacter pylori comprising: In a further embodiment, the invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl ) Amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof To a method of treating Chlamydia trachomatis.

  The term “treating” as used herein includes therapeutic and / or prophylactic treatment of the bacterial infections or other diseases described herein. This treatment includes the alleviation or alleviation of at least one symptom associated with a bacterial infection or at least one symptom associated with another disease described herein.

  As used herein, the term “therapeutically effective amount” is sufficiently high to alter the symptoms and / or condition to be treated fairly positively, within serious medical judgment, but with serious side effects. Means the amount of the compound or composition low enough (with a reasonable risk / effect ratio) to avoid The therapeutically effective amount of the active ingredient for use in the methods of the present invention is used for the particular disease being treated, the age and physical condition of the patient being treated, the severity of the disease, the duration of the treatment, the nature of the combination therapy, It will vary with the particular active ingredient, the particular pharmaceutically acceptable excipients utilized, and similar factors within the knowledge and experience of a skilled physician or veterinarian. A variety of suitable therapeutically effective amounts are described above.

  The term “subject” as used herein is an animal. “Subjects” include, but are not limited to, humans, mice, rats, guinea pigs, dogs, cats, horses, cows, pigs, monkeys, chimpanzees, baboons, or rhesus monkeys. In one embodiment, the “subject” is a mammal. In another embodiment, the “subject” is a human.

  As used herein, the phrase “pharmaceutically acceptable salt” of a compound means a salt that is pharmaceutically acceptable and that retains the desired pharmacological activity of the parent compound. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the compounds of the invention. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, mesylate, sulfate, bisulfate, phosphate , Acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, hydrogen tartrate, ascorbate, succinate, maleate, gentisate , Fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamo acid Salt (ie, 1,1′-methylene-bis- (2-hydroxy-3-naphthoate)). Suitable base salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. Preferably, the pharmaceutically acceptable salt is a crystalline salt. Even more preferably, the pharmaceutically acceptable salt is a crystalline salt selected from monohydrochloride, monomesylate, and monosulfate.

  Certain embodiments provide a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-Methicillin resistance including administration of crystalline monohydrochloride of octahydro-naphthacene-2-carboxylic acid amide It is directed to a method of treating S. aureus (MRSA). In this embodiment, the amount of monohydrochloride used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monohydrochloride is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monohydrochloride is administered daily.

  Another embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-methicillin resistance, including administration of crystalline monomesylate of octahydro-naphthacene-2-carboxylic acid amide It is directed to a method of treating S. aureus (MRSA). In this particular embodiment, the amount of crystalline monomesylate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monomesylate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monomesylate is administered daily.

  A further embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino)- Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide, including administration of crystalline monosulfate It is directed to a method of treating staphylococci (MRSA). In this particular embodiment, the amount of crystalline monosulfate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monosulfate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monosulfate is administered daily.

  Certain embodiments provide a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Administration of crystalline monohydrochloride of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide, Directed to a method of treating Trachomatis. In this embodiment, the amount of monohydrochloride used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monohydrochloride is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monohydrochloride is administered daily.

  Another embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Administration of a crystalline monomesylate of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Directed to a method of treating Trachomatis. In this particular embodiment, the amount of crystalline monomesylate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monomesylate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monomesylate is administered daily.

  A further embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino)- Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide crystalline monosulfate comprising administering Chlamydia trachomatis Directed to a method of treatment. In certain embodiments, the amount of crystalline monosulfate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monosulfate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monosulfate is administered daily.

  Certain embodiments provide a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Administration of crystalline monohydrochloride of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Directed to a method of treating H. pylori. In this embodiment, the amount of monohydrochloride used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monohydrochloride is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monohydrochloride is administered daily.

  Another embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Administering a crystalline monomesylate of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide, Directed to a method of treating H. pylori. In this particular embodiment, the amount of crystalline monomesylate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monomesylate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monomesylate is administered daily.

  A further embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino)- Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide crystalline monosulfate, comprising administration of Helicobacter pylori Directed to a method of treatment. In certain embodiments, the amount of crystalline monosulfate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monosulfate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monosulfate is administered daily.

  A further embodiment of the present invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) (Amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof Directed to a method for treating peptic ulcer. In a preferred embodiment, the method provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl Administering a crystalline salt of) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide. In this embodiment, the crystalline salt may be selected from the group consisting of monohydrochloride, monomesylate and monosulfate.

  Certain embodiments provide a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Digestion, including administering crystalline monohydrochloride of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Directed to a method of treating ulcers. In this embodiment, the amount of monohydrochloride used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monohydrochloride is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monohydrochloride is administered daily.

  Another embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Digestion, including administration of crystalline monomesylate of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Directed to a method of treating ulcers. In this particular embodiment, the amount of crystalline monomesylate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monomesylate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monomesylate is administered daily.

  A further embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino)- Peptic ulcer comprising administering crystalline monosulfate of methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Directed to a method of treatment. In certain embodiments, the amount of crystalline monosulfate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monosulfate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monosulfate is administered daily.

  In certain embodiments, the method of treating peptic ulcer further comprises administering to the subject at least one additional active ingredient, including but not limited to a proton pump inhibitor and / or bismuth. . A variety of suitable additional active ingredients for treating peptic ulcers are described in Bertram G. Katzung, “Basic and Clinical Pharmacology,” 1064-68 (8th edition, 2001), which is referred to in its entirety. Is incorporated herein by reference.

  A further embodiment of the present invention provides a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) (Amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof Directed to a method of treating Chlamydia pneumoniae. In a preferred embodiment, the method provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl Administering a crystalline salt of) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide. Preferably, the crystalline salt is selected from the group consisting of monohydrochloride, monomesylate and monosulfate.

  Certain embodiments provide a subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Administration of crystalline monohydrochloride of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide, Directed to a method of treating pneumoniae. In this embodiment, the amount of monohydrochloride used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monohydrochloride is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monohydrochloride is administered daily.

  Another embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) Administration of a crystalline monomesylate of -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Directed to a method of treating pneumoniae. In this particular embodiment, the amount of crystalline monomesylate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monomesylate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monomesylate is administered daily.

A further embodiment provides the subject with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino)- Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide crystalline monosulfate, comprising administering Chlamydia pneumoniae Directed to a method of treatment. In certain embodiments, the amount of crystalline monosulfate used is from about 10 mg to about 2000 mg, preferably from about 25 mg to about 500 mg. In certain embodiments, the monosulfate is administered at least once a month, preferably once a week, more preferably twice a week, and most preferably the monosulfate is administered daily.
Crystalline salt
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, 4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide and pharmaceutically acceptable salts thereof are used in the methods disclosed herein. In certain embodiments, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11- Dioxo-1,4,4a, 5,5a, 6,11,12a-crystalline monohydrochloride, crystalline monomesylate or crystalline monosulfate of octahydro-naphthacene-2-carboxylic acid amide is methicillin resistant Used to treat bacterial infections selected from the group consisting of S. aureus, Helicobacter pylori, Chlamydia trachomatis, and Chlamydia pneumoniae, or diseases associated with such infections.

  As used herein, the term “crystalline” refers to a solid state compound having a periodically repeating three-dimensional internal arrangement of atoms, ions or molecules characteristic of the crystal. The term crystalline does not necessarily mean that the compound exists as a crystal, but means that it has this crystal-like internal structure arrangement. As used herein, the term “amorphous” refers to a compound lacking a crystalline structure: no repetitive pattern, only short range order, extensive disorder.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Crystalline salt of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can be used to treat, prevent or ameliorate the following diseases: bacteria, Viruses, parasites, and fungal infections; cancer (eg, prostate, breast, colon, lung melanoma and lymphoma) and other diseases characterized by undesirable cell proliferation; arthritis; osteoporosis; diabetes; stroke; acute Myocardial infarction; aortic aneurysm; neurodegenerative diseases and other diseases where tetracycline compounds have been found to be active (see, eg, US Pat. Nos. 5,789,395; 5,834,450; 6,277,061; and 5,532,227) Each of which is incorporated herein by reference). Furthermore, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1, 4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide salt is a rickettsial infection, sexually transmitted infection, respiratory infection, bacterial infection, eye infection, anthrax Can be used to prevent or control important mammalian and veterinary diseases, such as; can serve as a treatment in acute intestinal amoebiasis, acne, Lyme disease, and peptic ulcers; Can be used for prevention.

  The term “monohydrochloride” as used herein refers to an ionic compound resulting from a neutralization reaction of an acid and a base. Since the ionic compound (here HCl) is composed of a cation and an anion, the compound is neutral.

  Common methods for analyzing crystalline salts include crystal analysis by powder X-ray diffraction (XRPD), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA).

The XRPD analysis disclosed herein was collected on a Bruker AXS C2 GADDS diffractometer using Cu Kα radiation (40 kV, 40 mA), an automatic XYZ stage, a laser video microscope for automatic sample positioning and a HiStar two-dimensional area detector. . The X-ray optical system consisted of a single Gobel multilayer mirror connected to a 0.3 mm pinhole collimator. The software used for data collection was GADDS for WNT 4.1.16, and the data were analyzed and presented using Diffrac Plus EVA v 9.0.0.2 or v 13.0.0.2. Samples were analyzed under ambient conditions as flat specimens using as received powder. About 1-2 mg of sample was lightly pressed on a glass slide to obtain a flat surface. Samples to be analyzed under non-ambient conditions were placed on a silicon wafer having a thermally conductive compound. The sample was then heated to the appropriate temperature at about 20 ° C. min ⁻¹ , followed by isothermal holding for about 1 minute before data collection was started.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11- The crystalline monohydrochloride of dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide is substantially shown in FIG. 1 after the synthesis of the crystalline salt It has such an XRPD pattern.

  As used herein, the term “XRPD pattern” refers to a graphical representation of the data collected by XRPD analysis. XRPD analysis is a technique used to characterize crystal structure, size, and preferred orientation in polycrystalline or powdered solid samples. This diffraction can also be used to characterize a heterogeneous solid mixture to determine the percent of crystalline compound present and can provide structural information about the unknown.

  The terms “substantially” and “about” as used herein with respect to an XRPD pattern are those where the described peak (s) appear within 0.2 degrees 2θ of a given 2θ value, for example within 0.1 degrees 2θ. Refers to the XRPD pattern.

  Crystalline monohydrochloride may have characteristic peaks that appear at least about 13.4, about 20.5, and about 23.3 at diffraction angles 2θ degrees as measured by XRPD. More preferably, the crystalline monohydrochloride has characteristic peaks appearing at least about 9.5, about 13.4, about 15.5, about 20.5, and about 23.3 at a diffraction angle 2θ degrees as measured by XRPD, and more preferably The crystalline monohydrochloride salt may have characteristic peaks appearing at a diffraction angle of 2θ degrees of at least about 9.5, about 13.4, about 15.5, about 16.6, about 19.2, about 20.5, about 22.2, and about 23.3.

  As used herein, the term “characteristic peak” refers to an XRPD pattern peak having an intensity that is at least 20%, more preferably 40% greater than baseline noise.

  TGA and DSC analysis are used to measure thermal behavior and can be used to distinguish polymorphs. One polymorphic form may exhibit different thermal behavior than an amorphous material or another polymorphic form.

The DSC analysis disclosed herein was collected on a TA Instruments Q2000 equipped with a 50 position autosampler. The instrument was calibrated for energy and temperature using certified indium. Calibration for heat capacity was performed using sapphire. Typically, 0.5-3.0 mg of each sample was heated from 25 ° C. to 250 ° C. at 10 ° C. min ⁻¹ in an aluminum pan with a pinhole. A nitrogen purge at 50 ml.min ^-1 was maintained on the sample. The instrument control software used was Advantage for Q Series v2.8.0.392 and Thermal Advantage v4.8.3, and the data was analyzed using Universal Analysis v4.4A.

  DSC is a thermal analysis technique that measures the difference in the amount of heat required to raise the temperature of a sample and a reference material as a function of temperature. DSC is used to measure some properties characteristic of a sample and allows observation of crystallization phenomena. In particular, DSC can be used to observe small energy changes that occur as a material transition from a solid to a liquid crystal and from a liquid crystal to an isotropic liquid. The presence of a phenomenon in the DSC curve can be used to assess the stability of the compound, as well as the presence of solvates or hydrates.

TGA is used to measure the change in weight associated with a change in temperature and can reveal the degradation of the compound and the presence of solvates or hydrates. The TGA analysis disclosed herein was collected on a TA Instruments Q500 TGA equipped with a 16 position autosampler. The instrument was calibrated for temperature using certified alumel and nickel. Typically, 5-30 mg of each sample was placed on a pre-weighed platinum crucible and aluminum DSC pan and heated from ambient to 300 ° C. at 10 ° C. min ⁻¹ . A nitrogen purge at 60 ml.min ^-1 was maintained on the sample. The instrument control and data analysis software used was Advantage for Q Series v2.8.0.392 and Thermal Advantage v4.8.3, and the data was analyzed using Universal Analysis v4.4A.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11- The crystalline monohydrochloride of dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide should exhibit a DSC curve substantially as shown in FIG. Preferably, it does not show any phenomenon until decomposition of the crystalline salt.

  As used herein, the term “phenomenon” refers to a change in a sample associated with heat absorption (endotherm) or generation (exotherm) that causes a change in differential heat flow recorded as a peak in a thermogram. Such changes in the sample include shape or form, solvate or hydrate degradation, degradation and changes. The absence of any phenomenon indicates that the compound is stable and in a low energy form.

  The term “substantially” as used herein with respect to a DSC curve means a DSC curve that exhibits a peak (s) within 1 ° C., eg, within 0.5 ° C. of a given temperature.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, The crystalline monohydrochloride of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can exhibit a TGA curve substantially as shown in FIG. About 1% to about 5% weight loss from about 30 ° C. to about 200 ° C., about 12% to about 16% weight loss from about 200 ° C. to about 250 ° C., more preferably from about 30 ° C. A weight loss of about 3% by about 200 ° C and a weight loss of about 14% to about 15% from about 200 ° C to about 250 ° C is exhibited.

  The term “substantially” as used herein with respect to a TGA curve means a curve that exhibits a weight loss percentage within 1%, eg, within 0.5%, of a predetermined value associated with a temperature change.

  The terms “stable” and “stability” as used herein refer to both the physical form and chemical purity of the salt. As used herein, “the salt” refers to the disclosed crystalline monohydrochloride, monomesylate and monosulfate salts of the present invention.

  As used herein, ambient conditions mean a temperature of about 20 ° C. to about 25 ° C. and about 40% RH.

  In another embodiment, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11- The crystalline monomesylate of dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can be used according to the method of the invention.

As used herein, the term “monomesylate” refers to an ionic compound resulting from a neutralization reaction of an acid and a base. Since this compound is composed of a cation and an anion (here CH ₃ SO ₂ ⁻ ), the compound is neutral.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, The crystalline monomesylate of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide has an XRPD pattern substantially as shown in FIG. 4 after synthesis of the crystalline salt .

  The crystalline monomesylate salt may have characteristic peaks that appear at least about 9, about 15, and about 23.8 at diffraction angles 2θ degrees as measured by XRPD. In preferred embodiments, the crystalline monomesylate has characteristic peaks appearing at at least about 9, about 15, about 22.7 and about 23.8 at diffraction angles 2θ degrees as measured by XRPD, more preferably, crystalline The sex monomesylate may have characteristic peaks appearing at least about 9, about 15, about 22, about 22.7 and about 23.8 at a diffraction angle of 2θ degrees.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, The crystalline monomesylate of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can exhibit a DSC curve substantially as shown in FIG. It is preferable that no phenomenon is exhibited until the decomposition of.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, The crystalline monomesylate of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can exhibit a TGA curve substantially as shown in FIG. About 1% to about 4% weight loss from about 30 ° C. to about 200 ° C., about 3% to about 10% weight loss from about 200 ° C. to about 250 ° C., more preferably from about 30 ° C. A weight loss of about 2% to about 3% by about 200 ° C and a weight loss of about 6% to about 7% from about 200 ° C to about 250 ° C is exhibited.

  In a further embodiment of the invention, (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1, Crystalline monosulfate of 11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can be used in the process of the present invention.

The term “monosulfate” as used herein refers to an ionic compound resulting from a neutralization reaction of an acid and a base. Since this compound is composed of a cation and an anion (here SO ₄ ²⁻ ), the compound is neutral.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Crystalline monosulfate of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide has an XRPD pattern substantially as shown in FIG. 7 after synthesis of the crystalline salt .

  Crystalline monosulfate may have characteristic peaks that appear at least about 15, about 17.8, and about 23.5 at diffraction angles 2θ degrees as measured by XRPD. In more preferred embodiments, the crystalline monosulfate salt may have characteristic peaks that appear at least about 15, about 17.8, about 22.5, and about 23.5 at diffraction angles 2θ degrees as measured by XRPD. In a more preferred embodiment, the crystalline monosulfate may have characteristic peaks that appear at at least about 15, about 17.8, about 19.0, about 22.5, and about 23.5 at diffraction angles 2θ degrees as measured by XRPD.

  (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Crystalline monosulfate of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can show a DSC curve substantially as shown in Figure 8 and analyzed by DSC It is preferable that the produced crystalline monosulfate does not show any phenomenon until the crystalline salt is decomposed.

(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Crystalline monosulfate of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide can exhibit a TGA curve substantially as shown in FIG. Crystalline monosulfate analyzed by TGA has a weight loss of about 1% to about 5% from about 30 ° C to about 200 ° C and a weight of about 12% to about 16% from about 200 ° C to about 250 ° C. A weight loss of about 3% to about 4% from about 30 ° C to about 200 ° C, and a weight loss of about 13% to about 14% from about 200 ° C to about 250 ° C.
Pharmaceutical composition One embodiment of the present invention is a bacterial infection selected from the group consisting of methicillin-resistant Staphylococcus aureus (MRSA, such as MRSA-CA and MRSA-HA), Helicobacter pylori, Chlamydia trachomatis, and Chlamydia pneumoniae Or (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12, for use in the treatment of infections or diseases associated with such infections (e.g. peptic ulcer and chlamydia) 12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide Or a pharmaceutically acceptable salt thereof and a pharmaceutical composition containing a pharmaceutically acceptable additive. Thus, in one aspect, the pharmaceutical composition of the present invention is used to treat peptic ulcer.

  In a preferred embodiment, the pharmaceutical composition comprises (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl]- Containing crystalline salt of 1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide, preferably monohydrochloride, monomesylate and It can be selected from the group consisting of monosulfates.

  The pharmaceutical composition of the present invention contains an effective amount of a crystalline salt, a pharmaceutically acceptable additive, and in some embodiments can further contain one or more additional active ingredients. . The content of the crystalline salt in the pharmaceutical composition of the present invention varies depending on, inter alia, the administration subject, the administration route and the target disease. The pharmaceutical composition of the present invention can be administered orally, topically (eg, transdermally, etc.), vaginally, rectally, or parenterally (eg, intravenously, etc.). Preferably, the pharmaceutical composition of the present invention is used to treat a bacterial infection.

  Examples of topical administration of the pharmaceutical composition include transdermal, buccal or sublingual application. For topical application, the pharmaceutical composition is preferably combined in a pharmacologically inert topical carrier such as a gel, ointment, lotion or cream. Such pharmacologically inert topical carriers include water, glycerol, alcohol, propylene glycol, fatty alcohol, triglyceride, fatty acid ester, or mineral oil. Other possible pharmacologically inert topical carriers are fluid petrolatum, isopropyl palmitate, polyethylene glycol, ethanol 95%, polyoxyethylene monolaurate 5% aqueous solution, sodium lauryl sulfate 5% aqueous solution, and the like. Furthermore, materials such as an antioxidant, a humectant, and a viscosity stabilizer may be added.

  For oral administration, the crystalline salts of the invention can be administered as capsules, tablets or granules. Tablets contain various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate and glycine, starch (preferably corn, potato or tapioca starch), alginic acid and certain complex silicates. It can be included with various disintegrants such as salts, together with granulating binders such as polyvinylpyrrolidone, sucrose, gelatin and gum arabic. In certain embodiments, the tablets may be film coated. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for tablets. Other solid compositions can also be used as fillers in gelatin capsules; preferred materials in this regard include lactose or lactose as well as high molecular weight polyethylene glycols. When aqueous suspensions and / or elixirs are desired for oral administration, the crystalline salt may be combined with various sweetening or flavoring agents, coloring materials or dyes, and optionally emulsified and / or suspended. Along with the agent, it can be combined with diluents such as water, ethanol, propylene glycol, glycerin and various mixtures thereof. The pharmaceutical composition of the present invention can be formulated such that the crystalline salt is released over a period of time after administration.

  Preparing such a pharmaceutical composition comprising a crystalline salt of the present invention, together with pharmaceutically acceptable excipients, and optionally additional active ingredients, may be any known in the art. This can be done by conventional techniques.

  In one embodiment, the crystalline salt present in the pharmaceutical composition is about 0.01 to about 90% by weight relative to the total composition. A suitable therapeutically effective amount of the crystalline salt will typically range from about 0.01 mg / kg to about 1 g / kg body weight / day; in another embodiment, from about 1 mg / kg to about 600 mg / kg. Body weight / day; in another embodiment, from about 1 mg / kg to about 250 mg / kg body weight / day; in another embodiment, from about 10 mg / kg to about 400 mg / kg body weight / day; in another embodiment, about 10 mg In another embodiment, from about 10 mg / kg to about 100 mg / kg body weight / day; in another embodiment, from about 10 mg / kg to about 25 mg / kg body weight / day; In another embodiment, from about 1 mg / kg to about 10 mg / kg body weight / day; in another embodiment, from about 0.001 mg / kg to about 100 mg / kg body weight / day; in another embodiment, from about 0.001 mg / kg to About 10 mg / kg body weight / day; and in another embodiment, about 0.001 mg / kg to about 1 mg / kg body weight / day. In certain embodiments, when the pharmaceutical composition described herein is administered orally, a suitable therapeutically effective amount of the crystalline salt is about 0.01 to about 100 mg / kg recipient body weight / day, preferably About 0.1 to about 50 mg / kg recipient body weight / day, more preferably about 0.1 to about 20 mg / kg recipient body weight / day, more preferably about 0.1 to about 10 mg / kg recipient body weight / day. Desirable doses can be administered once a day or in several divided doses, eg, 2-5 divided doses, at appropriate intervals throughout the day, or at other suitable schedules.

  As used herein, the term “pharmaceutically acceptable additive” includes, but is not limited to, one or more of the following: polymer, resin, plasticizer, filler, lubricant. , Diluents, binders, disintegrants, solvents, co-solvents, surfactants, buffer systems, preservatives, sweeteners, flavoring agents, pharmaceutical grade dyes or pigments, chelating agents, viscosity modifiers, and combinations thereof. Pharmaceutically acceptable additives can be used in any component in making the dosage form, ie, the core tablet or coating. Particularly useful flavoring agents and dyes and pigments herein include, but are not limited to, those described in the Handbook of Pharmaceutical Excipients (4th Edition, Pharmaceutical Press 2003). Suitable co-solvents include, but are not limited to, ethanol, isopropanol, acetone, and combinations thereof. Suitable surfactants include, but are not limited to, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene monoalkyl ethers, sucrose monoesters, simethicone emulsions, sodium lauryl sulfate, Tween 80®, and lanolin. Examples include esters, ethers, and combinations thereof. Suitable preservatives include, but are not limited to, phenol, alkyl esters of parahydroxybenzoic acid, benzoic acid and its salts, boric acid and its salts, sorbic acid and its salts, chlorbutanol, benzyl alcohol, thimerosal, Examples include acetic acid and phenylmercuric nitrate, nitromerzole, benzalkonium chloride, cetylpyridinium chloride, methylparaben, propylparaben, and combinations thereof. Suitable fillers include but are not limited to starch, lactose, sucrose, maltodextrin, and microcrystalline cellulose. Suitable plasticizers include, but are not limited to, triethyl citrate, polyethylene glycol, propylene glycol, dibutyl phthalate, castor oil, acetylated monoglycerides, triacetin, and combinations thereof. Suitable polymers include, but are not limited to, ethyl cellulose, cellulose acetate trimellitate, hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate, polyvinyl acetate phthalate, Eudragit® L 30-D, Eudragit® L 100-55, Eudragit® F530D and Eudragit® S 100 (Rohm Pharma GmbH and Co. KG, Darmstadt, Germany), Acryl-EZE® and Sureteric® (Colorcon, West Point, Pennsylvania), and combinations thereof. Suitable lubricants include, but are not limited to, magnesium stearate, stearic acid, talc, and combinations thereof.

  As used herein, the term “additional active ingredient” includes any known in the art to treat, prevent, or alleviate a symptom treated by the pharmaceutical composition. Of drugs. Such agents include, but are not limited to, agents that are known to treat, prevent, or reduce the symptoms of bacterial infections and inflammatory skin diseases. In one embodiment where the pharmaceutical composition of the present invention is used to treat peptic ulcers, additional active ingredients include, but are not limited to, proton pump inhibitors and / or bismuth. Additional agents for treating peptic ulcers are described in Bertram G. Katzung, “Basic and Clinical Pharmacology,” 1064-68 (8th edition, 2001), which is incorporated herein by reference in its entirety. It is done.

  In certain embodiments, the pharmaceutical composition comprises a bacterial infection, such as methicillin-resistant Staphylococcus aureus (MRSA, such as MRSA-CA and MRSA-HA), Helicobacter pylori, Chlamydia trachomatis, Chlamydia pneumoniae, or such (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7 for use in the treatment of diseases associated with infection (e.g. peptic ulcer and chlamydia) -[(Methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide monohydrochloride and pharmacy Containing additives that are acceptable to the environment. In another embodiment, the pharmaceutical composition comprises a bacterial infection, such as methicillin-resistant Staphylococcus aureus (MRSA, such as MRSA-CA and MRSA-HA), Helicobacter pylori, Chlamydia trachomatis, Chlamydia pneumoniae, or such (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7 for use in the treatment of diseases associated with infection (e.g. peptic ulcer and chlamydia) -[(Methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid monomesylate salt and pharmacy Containing additives that are acceptable to the environment. In further embodiments, the pharmaceutical composition comprises a bacterial infection, such as methicillin-resistant Staphylococcus aureus (MRSA, such as MRSA-CA and MRSA-HA), Helicobacter pylori, Chlamydia trachomatis, Chlamydia pneumoniae, or such an infection. (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-, for use in the treatment of diseases related to infectious diseases (e.g. peptic ulcer and chlamydia) [(Methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid monosulfate and pharmaceutical Contains acceptable additives.

  The following examples illustrate the practice of this invention in some of the preferred embodiments. Other embodiments within the scope of the claims will be apparent to those skilled in the art.

The following examples illustrate the synthesis of the compounds described herein.
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Synthesis of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide ("free base")

N,N-ジメチルアセトアミド(15mL)中の7-ホルミルサンサイクリンTFA塩(2.23g)とN,O-ジメチルヒドロキシルアミン塩酸塩(780mg)の溶液を、アルゴン雰囲気下に室温で10分間撹拌した。この溶液にシアノ水素化ホウ素ナトリウム(302mg)を添加した。この溶液を5分間撹拌して、LC-MSによってモニタリングした。反応混合物をジエチルエーテルの中に注ぎ、生じた沈殿物を真空下でろ過により集めた。この粗生成物を、C18カラム(20mMトリエタノールアミン水溶液中の10〜40％アセトニトリルの直線勾配、pH7.4)を用いた分取HPLCにより精製した。分取HPLCの画分を集めて、有機溶媒(アセトニトリル)を減圧下で蒸発させた。得られた水性溶液をクリーンなPDVB SPEカラムにロードし、蒸留水で洗浄し、次に0.1M酢酸ナトリウム溶液、続いて蒸留水で洗浄した。生成物をアセトニトリルで溶出させた。溶出液を減圧下で濃縮すると、385mgが遊離塩基として得られた。
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸の結晶性モノ塩酸塩(「結晶性モノ塩酸塩」)の合成

A solution of 7-formyl sancycline TFA salt (2.23 g) and N, O-dimethylhydroxylamine hydrochloride (780 mg) in N, N-dimethylacetamide (15 mL) was stirred at room temperature for 10 minutes under an argon atmosphere. To this solution was added sodium cyanoborohydride (302 mg). The solution was stirred for 5 minutes and monitored by LC-MS. The reaction mixture was poured into diethyl ether and the resulting precipitate was collected by filtration under vacuum. The crude product was purified by preparative HPLC using a C18 column (linear gradient of 10-40% acetonitrile in 20 mM aqueous triethanolamine, pH 7.4). Preparative HPLC fractions were collected and the organic solvent (acetonitrile) was evaporated under reduced pressure. The resulting aqueous solution was loaded onto a clean PDVB SPE column, washed with distilled water, then washed with 0.1 M sodium acetate solution followed by distilled water. The product was eluted with acetonitrile. The eluate was concentrated under reduced pressure to give 385 mg as the free base.
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Synthesis of crystalline monohydrochloride of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid ("crystalline monohydrochloride")

粗製の(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミド(100g, 含量約35％)を分取カラムクロマトグラフィーで精製した。目的の画分(8〜10リットル)を一緒に合わせて、水酸化アンモニウムを用いてpHを7.0〜7.5に調整した。この水性溶液をジクロロメタンで3回抽出した(各回4リットル)。ジクロロメタン相を一緒に合わせて、減圧下で濃縮した。残留物をエタノール(800ml)中に懸濁させ、20mlの水を加えた。メタノール中の1.25M塩酸を用いてpHをpH1.6〜1.3に徐々に調整し、その混合物を20〜60分間撹拌した。この時点で遊離塩基は完全に溶解した。その溶液を200〜250mlへと減圧下で濃縮し、(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミド・モノHCl結晶(100〜200mg)を種晶として加えた。スラリーを≦5℃に保ちながら、撹拌を2〜18時間続けた。得られた結晶をろ過し、エタノール(50mL)で洗浄し、一定の重量になるまで減圧下で乾燥させた。20gの結晶性(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミド・モノ塩酸塩が純度≧90％、含量≧90％で単離された。
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸の結晶性モノメシル酸塩(「結晶性メシル酸塩」)の合成
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミド遊離塩基(74mg)をエタノール(740μl)中に懸濁させ、撹拌しながら60℃(浴温度)に加熱した。メタンスルホン酸(1.1当量, 1M THF溶液として167μl)を加えたところ、固体のほとんどが溶解した。5分後、この懸濁液を(油浴で制御しないで)約1.75時間かけて周囲温度にまで冷やした。53℃までに固体が沈殿し、これを減圧下に周囲温度でろ過した。この懸濁液は粘性であったので、ろ過を助けるために追加のエタノール(200μl)を加えた。ケーキをn-ヘキサン(400μl)で洗浄し、フィルター上で約30分間空気乾燥させると、59mg(収率67％)の黄色の固体が得られた。
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸の結晶性モノ硫酸塩(「結晶性硫酸塩」)の合成
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミド遊離塩基(86mg)をエタノール(500μl)中に懸濁させ、撹拌しながら63℃(浴温度)に加熱した。この温度で遊離塩基の大部分が溶解した。硫酸(1.1当量, 1M水溶液として194μl)を加えたところ、全部の固体が溶解した。この溶液を(油浴で制御しないで)約1.75時間かけて周囲温度にまで冷やした。この温度では固体はまったく析出してこなかった。メチルt-ブチルエーテル(MtBE)を貧溶媒(4×50μl)として添加した。各添加は曇り点を生じさせたが、撹拌すると固体は再溶解した。この溶液をストッパーで約3時間撹拌した後に固体が析出した。その固体を減圧下にろ過し、MtBE (3×200μl)で洗浄し、フィルター上で約45分間空気乾燥させると、93mg(収率90％)の黄色の固体が得られた。
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミドの非晶質ビス塩酸塩の合成
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミド遊離塩基(1g)をメタノール(50mL)中に懸濁させた。過剰のメタノール性HClを加えることによって遊離塩基を塩酸塩に変換し、続いて減圧下で蒸発させたところ、1.1gの黄色の固体が得られた：MS (Mz+1 = 488). 1H NMR (300 MHz, CD3OD) δ 7.46 (d, 1H, J = 8.6 Hz), 6.81 (d, 1H, J = 8.6 Hz), 4.09 (d, 1H, J = 1.0 Hz), 3.79 (d, 1H, J = 13.1 Hz), 3.73 (d, 1H, J = 13.1 Hz), 3.36 (m, 1H), 3.27 (s, 3H), 3.08-2.95 (8H), 2.61 (s, 3H), 2.38 (t, 1H, J = 14.8), 2.22 (m, 1H), 1.64 (m, 1H)。XRPDパターンは図10に示されており、TGAおよびDSC曲線は重ね合わせて図11に示されている。
(4S,4aS,5aR,12aS)-4-ジメチルアミノ-3,10,12,12a-テトラヒドロキシ-7-[(メトキシ(メチル)アミノ)-メチル]-1,11-ジオキソ-1,4,4a,5,5a,6,11,12a-オクタヒドロ-ナフタセン-2-カルボン酸アミドの非晶質モノ塩酸塩の合成
結晶性モノ塩酸塩のサンプル(2.09g)を水(250ml, 120倍容)に溶解し、ろ過し、-78℃浴中で凍結させた。凍結乾燥機を用いて、固化したサンプルから水を110時間かけて除去すると、非晶質のモノ塩酸塩がフワフワした黄色の固体として得られ、これはXRPD解析により非晶質であることが確認された。
抗菌活性試験
結晶性モノ塩酸塩の抗菌活性は、本明細書に詳述される抗嫌気性菌活性および作用機序試験に従って評価した。

Crude (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1, 4,4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide (100 g, content about 35%) was purified by preparative column chromatography. The desired fractions (8-10 liters) were combined together and the pH was adjusted to 7.0-7.5 using ammonium hydroxide. This aqueous solution was extracted three times with dichloromethane (4 liters each time). The dichloromethane phases were combined together and concentrated under reduced pressure. The residue was suspended in ethanol (800 ml) and 20 ml water was added. The pH was gradually adjusted to pH 1.6-1.3 using 1.25M hydrochloric acid in methanol and the mixture was stirred for 20-60 minutes. At this point, the free base was completely dissolved. The solution was concentrated to 200-250 ml under reduced pressure and (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -Methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide mono HCl crystals (100-200 mg) were added as seed crystals . Stirring was continued for 2-18 hours while maintaining the slurry at ≦ 5 ° C. The obtained crystals were filtered, washed with ethanol (50 mL), and dried under reduced pressure until a constant weight was reached. 20 g of crystalline (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo- 1,4,4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide monohydrochloride was isolated with a purity ≧ 90% and content ≧ 90%.
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Synthesis of crystalline monomesylates of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid ("crystalline mesylate")
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, 4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide free base (74 mg) was suspended in ethanol (740 μl) and heated to 60 ° C. (bath temperature) with stirring. Methanesulfonic acid (1.1 eq, 167 μl as 1M THF solution) was added and most of the solid dissolved. After 5 minutes, the suspension was cooled to ambient temperature over approximately 1.75 hours (not controlled by an oil bath). A solid precipitated by 53 ° C. and was filtered at ambient temperature under reduced pressure. Since this suspension was viscous, additional ethanol (200 μl) was added to aid filtration. The cake was washed with n-hexane (400 μl) and air dried on the filter for about 30 minutes to give 59 mg (67% yield) of a yellow solid.
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Synthesis of crystalline monosulfate ("crystalline sulfate") of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, 4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide free base (86 mg) was suspended in ethanol (500 μl) and heated to 63 ° C. (bath temperature) with stirring. Most of the free base dissolved at this temperature. Sulfuric acid (1.1 eq, 194 μl as a 1M aqueous solution) was added and all solids dissolved. The solution was cooled to ambient temperature over approximately 1.75 hours (not controlled by an oil bath). At this temperature, no solid was deposited. Methyl t-butyl ether (MtBE) was added as an anti-solvent (4 × 50 μl). Each addition produced a cloud point, but the solid redissolved when stirred. After this solution was stirred with a stopper for about 3 hours, a solid precipitated. The solid was filtered under reduced pressure, washed with MtBE (3 × 200 μl) and air dried on the filter for about 45 minutes to give 93 mg (90% yield) of a yellow solid.
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Synthesis of amorphous bishydrochloride of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, 4a, 5,5a, 6,11,12a-Octahydro-naphthacene-2-carboxylic acid amide free base (1 g) was suspended in methanol (50 mL). The free base was converted to the hydrochloride salt by adding excess methanolic HCl, followed by evaporation under reduced pressure to give 1.1 g of a yellow solid: MS (Mz + 1 = 488). 1H NMR (300 MHz, CD3OD) δ 7.46 (d, 1H, J = 8.6 Hz), 6.81 (d, 1H, J = 8.6 Hz), 4.09 (d, 1H, J = 1.0 Hz), 3.79 (d, 1H, J = 13.1 Hz), 3.73 (d, 1H, J = 13.1 Hz), 3.36 (m, 1H), 3.27 (s, 3H), 3.08-2.95 (8H), 2.61 (s, 3H), 2.38 (t, 1H , J = 14.8), 2.22 (m, 1H), 1.64 (m, 1H). The XRPD pattern is shown in FIG. 10, and the TGA and DSC curves are shown superimposed in FIG.
(4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4, Synthesis of amorphous monohydrochloride of 4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide A sample of crystalline monohydrochloride (2.09 g) was added to water (250 ml, 120 volumes) , Filtered and frozen in a -78 ° C bath. When water is removed from the solidified sample over 110 hours using a freeze dryer, amorphous monohydrochloride is obtained as a fluffy yellow solid, which is confirmed to be amorphous by XRPD analysis It was done.
Antibacterial Activity Test The antibacterial activity of crystalline monohydrochloride was evaluated according to the anti-anaerobic activity and mechanism of action tests detailed herein.

  In the tests below, the samples were prepared using bishydrochloride and the data are expressed based on the free base (“active substance”). The overall anti-anaerobic activity of crystalline monohydrochloride can be seen from in vitro testing of the active substance against 37 representative anaerobic strains and the results are summarized in Table 1. The active substance showed relatively strong activity (i.e. minimum inhibitory concentration (MIC) of 4 μg / mL or less) against many species of Gram-positive bacteria, including P. acnes . Overall, the activity of the active substance was similar to that of tetracycline and doxycycline, but inferior to that of minocycline. Organisms that showed high MIC values (MIC ≧ 16 μg / mL) for the active substance include C. perfringens and S. constellatus.

  The MIC values for gram negative anaerobes are shown in Table 1. Tetracycline resistant strains showed cross resistance to the active substance. Active substances and other tetracyclines are potent activity against E. corrodens and Fusobacterium spp., P. melaninogenica (one of two strains) and It showed moderate activity against V. parvula and weak activity against P. asaccharolytica.

活性の抗菌スペクトル
活性物質の活性の抗菌スペクトルの評価は、種々のグラム陽性およびグラム陰性の好気性および嫌気性生物についてのin vitro MIC測定により、いくつかの試験で判定された。これらのアッセイの結果(表2に要約)から、活性物質は臨床的に使用されるテトラサイクリン類よりも狭い活性スペクトルで、プロピオン酸菌と他のグラム陽性菌に対する活性を明示したことが示される。テトラサイクリンに耐性の菌株は、活性物質に交差耐性を示す。各生物グループに対する活性は、表の後の本文中で説明される。

Evaluation of antibacterial spectrum of activity of the active antibacterial spectrum actives by in vitro MIC measurement for aerobic and anaerobic organisms of various gram-positive and gram-negative, is determined by several tests. The results of these assays (summarized in Table 2) show that the active substance has demonstrated activity against propionic acid bacteria and other gram-positive bacteria with a narrower spectrum of activity than clinically used tetracyclines. Strains resistant to tetracycline exhibit cross resistance to the active substance. The activity against each organism group is explained in the text after the table.

クラミジア・トラコマチスに対する活性
下記の試験では、サンプルは結晶性モノ塩酸塩を用いて調製したものであり、データは遊離塩基(「活性物質」)に基づいて表されている。クラミジア・トラコマチスに対する該化合物のin vitro活性を、アジスロマイシン、レボフロキサシン、およびドキシサイクリンのそれと比較した。in vitro感受性試験はHEp-2細胞で実施した。C.トラコマチスの菌株の90％が阻止される活性物質のMIC(MIC₉₀)は、0.125μg/mL(範囲0.06〜0.125μg/mL)であった。菌株の90％が阻止される最小殺菌濃度(MBC₉₀)も0.125μg/mLであった。これらのデータは、比較薬剤から得られたデータと類似していた(アジスロマイシン、レボフロキサシンおよびドキシサイクリンのMIC₉₀は、それぞれ0.015、0.25および0.125μg/mLであった)。MICは、試験した分離株の地理的分布に関係なく、分離株間で非常に一致していた。
ヘリコバクター・ピロリ(H.ピロリ)に対する活性
下記の試験では、サンプルは結晶性モノ塩酸塩を用いて調製したものであり、データは遊離塩基(「活性物質」)に基づいて表されている。H.ピロリのさまざまな菌株に対する該化合物のin vitro活性をアモキシシリン、テトラサイクリンおよびメトロニダゾールのそれと比較した(表3参照)。活性物質は、試験した菌株に対して一致した活性を示し、すべての試験菌株を8μg/mL以下で阻止し、13のうち12の菌株は1〜2μg/mLで阻止された。臨床分離株については、MIC₉₀値は2μg/mLであった。活性物質は3種の比較薬剤よりも一般的に低い活性であった；しかしながら、活性のレベルは、経口投与後に上部消化管で達成され得る薬物レベルを考慮すると、顕著である。

Activity against Chlamydia trachomatis In the following test, the sample was prepared using crystalline monohydrochloride and the data is expressed on the free base ("active substance"). The in vitro activity of the compound against Chlamydia trachomatis was compared with that of azithromycin, levofloxacin, and doxycycline. In vitro susceptibility testing was performed on HEp-2 cells. The MIC (MIC ₉₀ ) of the active substance at which 90% of C. trachomatis strains are blocked was 0.125 μg / mL (range 0.06-0.125 μg / mL). The minimum bactericidal concentration (MBC ₉₀ ) at which 90% of the strains were blocked was also 0.125 μg / mL. These data were similar to those obtained from the comparative drug (the MIC _{90 for} azithromycin, levofloxacin and doxycycline were 0.015, 0.25 and 0.125 μg / mL, respectively). The MIC was very consistent between isolates regardless of the geographical distribution of isolates tested.
Activity against Helicobacter pylori (H. pylori) In the tests below, samples were prepared using crystalline monohydrochloride, and data are expressed based on the free base ("active substance"). The in vitro activity of the compound against various strains of H. pylori was compared to that of amoxicillin, tetracycline and metronidazole (see Table 3). The active substances showed consistent activity against the tested strains, blocking all tested strains at 8 μg / mL or less, and 12 strains out of 13 being blocked at 1-2 μg / mL. For clinical isolates, the MIC ₉₀ value was 2 μg / mL. The active substance was generally less active than the three comparative drugs; however, the level of activity is significant considering the drug levels that can be achieved in the upper gastrointestinal tract after oral administration.

メチシリン耐性黄色ブドウ球菌(MRSA)に対する活性
下記の試験では、サンプルは結晶性モノ塩酸塩を用いて調製したものであり、データは遊離塩基(「活性物質」)に基づいて表されている。メチシリン感受性黄色ブドウ球菌(MSSA)、院内感染型メチシリン耐性黄色ブドウ球菌(MRSA-HA)または市中感染型メチシリン耐性黄色ブドウ球菌(MRSA-CA)のさまざまな菌株に対する該化合物のin vitro活性を、MRSAの治療によく用いられる経口抗菌剤であるドキシサイクリン、オキサシリン、クリンダマイシン、リネゾリド、およびトリメトプリム/スルファメトキサゾール(TMP/SXT)の組み合わせのそれと比較した。MICの結果を表4にまとめてある。活性物質はMSSAに対して非常に活性であり、0.25〜16μg/mLのMIC範囲、0.25μg/mLのMIC₅₀、および1μg/mLのMIC₉₀を示した；これらの値はドキシサイクリンおよびオキサシリンのそれに類似していた。ドキシサイクリン耐性MSSAの単一の菌株(8μg/mLのDOX MIC)はまた、活性物質に対しても上昇したMIC(16μg/mL)を有していた。活性物質は、MSSAに対してTMP/SXTよりも活性が低かったが、クリンダマイシンまたはリネゾリドよりも活性であった。試験生物のすべてがリネゾリドに感受性であった。

Activity Against Methicillin Resistant Staphylococcus aureus (MRSA) In the tests below, samples were prepared using crystalline monohydrochloride and data are expressed based on the free base ("active substance"). In vitro activity of the compound against various strains of methicillin-sensitive Staphylococcus aureus (MSSA), hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA-HA) or community-acquired methicillin-resistant Staphylococcus aureus (MRSA-CA), Compared to that of the combination of doxycycline, oxacillin, clindamycin, linezolid, and trimethoprim / sulfamethoxazole (TMP / SXT), which are commonly used oral antimicrobial agents for the treatment of MRSA. Table 4 summarizes the MIC results. The active substance was very active against MSSA and showed a MIC range of 0.25-16 μg / mL, a MIC _{50 of} 0.25 μg / mL, and a MIC _{90 of} 1 μg / mL; these values are that of doxycycline and oxacillin It was similar. A single strain of doxycycline resistant MSSA (8 μg / mL DOX MIC) also had an elevated MIC (16 μg / mL) for the active substance. The active substance was less active against MSSA than TMP / SXT, but more active than clindamycin or linezolid. All of the test organisms were sensitive to linezolid.

The active substance showed a potent activity against MRSA-HA, equivalent to doxycycline, with MIC ₅₀ and MIC ₉₀ values of 0.25 μg / mL and 0.5 μg / mL, respectively. The active substance was less active than TMP / SXT and more active than the remaining comparative agents. All of the organisms were sensitive to linezolid.

Finally, the active agent shows a strong activity against MRSA-CA, it had MIC ₉₀ values of 0.5 [mu] g / mL which is similar to the MIC ₉₀ of doxycycline and clindamycin. A single strain of doxycycline resistant MRSA-CA (8 μg / mL DOX MIC) also had an elevated MIC (8 μg / mL) for the active substance. The active substance was less active than TMP / SXT, but more active than oxacillin and linezolid. All of the test organisms were sensitive to linezolid.

クラミジア・トラコマチスおよびクラミジア・ニューモニエに対する活性
この試験では、サンプルは結晶性モノ塩酸塩を用いて調製したものであり、データは遊離塩基(「活性物質」)に基づいて表されている。クラミジア・トラコマチス(C.トラコマチスともいう)およびクラミジア・ニューモニエ(C.ニューモニエともいう)の10分離株に対する活性物質のin vitro活性を試験して、3種の比較薬剤：レボフロキサシン、アジスロマイシンおよびドキシサイクリンの活性と比較した。

Activity against Chlamydia trachomatis and Chlamydia pneumoniae In this test, samples were prepared using crystalline monohydrochloride and data are expressed based on the free base ("active substance"). The in vitro activity of the active substance against 10 isolates of Chlamydia trachomatis (also called C. tramonitis) and Chlamydia pneumoniae (also called C. pneumoniae) was tested and the activity of three comparative drugs: levofloxacin, azithromycin and doxycycline Compared with.

The following materials were used in this test:
1. Chlamydia isolates • C. trachomatis isolates include standard isolates from ATCC®: D-UW-57Cx (VR-878), E-BOUR (VR-348B), F-IC- CAL3 (VR-346), H-UW-43Cx (VR-879), I-UW-12Ur (VR-880), J-UW-36Cx (VR-886), L2-434 (VR-902B) and clinical Isolates N18 (cervical), N19 (cervical), 7015 (infant eyes) were included.

      • C. pneumoniae isolates tested included five isolates derived from standard isolates from ATCC: TW 183, AR 39 (53592), CM-1 (VR-1360), T 2040, and US human immunity Six isolates (BAL15, BAL16, BAL 18, BAL 19, BAL 37, BAL 62) from bronchoalveolar lavage samples obtained from patients with deficiency virus and pneumonia were included.

2. HEp2 cells: ATCC® [Manassas, VA] ATCC® number: CCL-23 ™ Lot # 58978772
3. Antibiotics
a. Active substance
b. Three comparative drugs manufactured by Sigma-Aldrich (registered trademark)
i. Levofloxacin Catalog No. 28266-1G-F Lot # BCBF7004V
ii. Doxycycline Catalog No. D9891-1G Lot # BCBD5187V
iii. Azithromycin Cat # 75199-25MG-F Lot # E446421 / 1V
4. IMDM with L-glutamine and phenol red: Gibco® catalog number 12440-061 Lot # 1153614
5. FBS: Gibco (R) catalog number 10438-026 Lot # 1140649
6. Cycloheximide: Sigma-Aldrich® catalog number C4859 lot # 090M4009
7. PBS: Gibco® catalog number 10010-072 Gibco lot # 1144930
8. Pathfinder® Chlamydia Culture Confirmation System: Biorad® catalog number 30701 Lot # 109515

The active substances, azithromycin, levofloxacin, and doxycycline were provided as powders and solubilized according to the manufacturer's instructions. A 1280 μg / ml stock solution was made and frozen at −80 ° C. Aliquots of stock drug suspension were diluted each time the assay was performed. Chlamydia isolates were grown to a concentration of 10 ⁷ to 10 ⁸ inclusion-forming units (IFU) / ml by continuous passage in tissue culture using a medium without antibiotics. The isolate was purified by centrifuging at 500 rpm to sediment cell debris. Chlamydia containing supernatant was pelleted at 17,000 xg for 1 hour. The pellet containing Chlamydia was then resuspended in sucrose-phosphate-glutamate medium (SPG) and centrifuged through a discontinuous renographin gradient. SPG is composed of sucrose (250 mM), glutamic acid (5 mM), sodium phosphate (10 mM), and 20% fetal calf serum and has a pH of 7.4. The band containing Chlamydia elementary body (EB) was then washed 3 times and resuspended in SPG. The EB suspension is titrated in HEp-2 cells.

Sensitivity tests for C. pneumoniae and C. trachomatis were performed in cell culture using HEp-2 cells grown in 96-well microtiter plates. Each well was inoculated with 0.2 ml of the test strain diluted to 10 ⁴ inclusion body forming units / ml; the plate was centrifuged at 1,700 × g for 1 hour and incubated at 35 ° C. for 1 hour. The wells were then aspirated and overlaid with a medium containing 1 μg / ml cycloheximide and a serial 2-fold dilution of the test drug. After incubating at 35 ° C. for 72 hours, the cultures were fixed and stained for inclusion bodies using a fluorescein-conjugated antibody against the genus-specific antigen of Chlamydia lipopolysaccharide (Pathfinder; Biorad, Redmond, WA). The minimum inhibitory concentration (MIC) was the lowest antibiotic concentration at which no inclusion bodies were seen. Minimum bactericidal concentration (MBC) was determined by aspirating antibiotic-containing medium, washing the wells twice with phosphate buffered saline, and adding antibiotic-free medium. Infected cells were frozen at -70 ° C, thawed, passaged to new cells, incubated for 72 hours, then fixed and stained as described above. MBC was the lowest antibiotic concentration that did not allow inclusion bodies to form after passage. All studies were performed in duplicate (Roblin, PM et al., “In vitro activity of CEM-101, a new fluoroketolide antibiotic, against Chlamydia trachomatis and Chlamydia (Chlamydophila) pneumonia,” Antimicrob Agents Chemother., Vol. 54, No. 3, pp. 1358-1359 (2010)). The MIC and MIB results for C. trachomatis and C. pneumoniae in this study are shown in Tables 5 and 6.

活性物質について、C.トラコマチスの分離株の90％が阻止されたMICは、0.125μg/ml(範囲0.03〜0.125μg/ml)であった。C.ニューモニエの分離株のMIC₉₀は、0.5μg/ml(範囲0.125〜0.5μg/ml)であった。分離株の90％が活性物質によって死滅した最小殺菌濃度(MBC₉₀)は、C.トラコマチスについて0.125μg/ml(範囲0.03〜0.125μg/ml)であり、C.ニューモニエについて0.25μg/ml(範囲0.125〜0.5μg/ml)であった。レボフロキサシン、ドキシサイクリン、およびアジスロマイシンのMBC₉₀は、それぞれ2、0.5、および0.25μg/mlであった。

For the active substance, the MIC at which 90% of C. trachomatis isolates were blocked was 0.125 μg / ml (range 0.03-0.125 μg / ml). The MIC ₉₀ of the C. pneumoniae isolate was 0.5 μg / ml (range 0.125 to 0.5 μg / ml). The minimum bactericidal concentration (MBC ₉₀ ) at which 90% of the isolates were killed by the active substance is 0.125 μg / ml (range 0.03-0.125 μg / ml) for C. trachomatis and 0.25 μg / ml (range) for C. pneumoniae 0.125 to 0.5 μg / ml). The MBC _{90 for} levofloxacin, doxycycline, and azithromycin was 2, 0.5, and 0.25 μg / ml, respectively.

These results therefore show that the in vitro activity of the active substance against C. trachomatis and C. pneumoniae was comparable to doxycycline.
Mechanism of Action In the tests below, samples were prepared using bishydrochloride and data are expressed based on the free base (“active substance”). The mechanism of action of crystalline monohydrochloride was determined by two different approaches through active substance testing, as described below.

In the first approach, “Antimicrobial Action Mechanism: In Vitro Inhibition of Bacterial Transcription and Translation,” the ability of active substances to inhibit bacterial protein synthesis is demonstrated by an in vitro cell-free bacterial transcription and translation assay (Promega (Madison (Beckler, G., Promega Notes 31 (1991) pp. 3-6). The active substance inhibited the synthesis of the reporter protein with an IC ₅₀ of 8.3 ± 0.18 μM. This value was comparable to the measured IC ₅₀ values for comparison drug tetracycline, doxycycline and minocycline (IC ₅₀ values of 4.7 ± 0.48 and 2.4 ± 0.22 [mu] M). These results provide evidence that the active substance functions as a classic tetracycline by inhibiting bacterial protein synthesis.

In the second approach, “Antimicrobial Action Mechanism: Inhibition of Macromolecular Synthesis in Staphylococcus aureus”, the ability of the active substance to target bacterial protein synthesis is based on macromolecular synthesis in Gram-positive bacteria, Staphylococcus aureus. Further confirmation in whole cell assays. The active substance inhibited the uptake of [ ³ H] -leucine into the protein of the growing organism within a concentration range of 0.25 to 8 times the MIC (0.063 to 2 μg / mL) in a dose-dependent manner. A maximum inhibition of 80% was observed at 8 times the MIC, which was comparable to that observed for the tetracycline comparison drugs doxycycline and minocycline. In contrast, 8 times the active substance of the MIC showed less than 20% inhibition of the synthesis of the cell wall, DNA, RNA and lipid components of the test bacteria. The results of this test indicate that the active substance acts as a selective inhibitor of bacterial protein synthesis at concentrations comparable to known tetracyclines.

  The above in vitro susceptibility tests included tetracycline resistant strains with characterized tetracycline resistant genes. Strains are the most commonly found tetracycline resistance genes: those that carry efflux (tetK, tet38, tetL, tetS, tetB, and tetD), ribosome protection (tetM and tetO), and resistant by rRNA point mutations P. Acnes was selected. The MIC values for these selected strains showed some cross resistance between the active substance and other tetracyclines as shown in Table 7. The presence of the tetracycline resistance gene increased the MIC of the active substance compared to susceptible strains (except for tetK in S. aureus), which MIC values were similar to those of doxycycline and / or minocycline, but Generally lower than that.

本明細書に開示された好ましい実施形態の多数の変更、修飾、および変形は当業者には明らかであろう。それらはすべて、特許請求された本発明の精神および範囲内であると予想され、かつ考えられる。例えば、特定の実施形態が詳細に説明されているが、当業者であれば、前述の実施形態および変形形態はさまざまなタイプの代用の、追加の、または代替の材料を組み込むために改変され得ることが理解されよう。したがって、本発明のほんのわずかな変形形態しか本明細書に記載されていないとしても、そのような追加の変更および変形形態ならびにそれらの均等形態の実施は、以下の特許請求の範囲に定義される本発明の精神および範囲内にあることが理解されるべきである。本明細書に引用された全ての特許出願、特許および他の刊行物は、その全体が参照により本明細書に組み入れられる。

Numerous changes, modifications, and variations of the preferred embodiments disclosed herein will be apparent to those skilled in the art. All are anticipated and considered to be within the spirit and scope of the claimed invention. For example, although specific embodiments have been described in detail, those of ordinary skill in the art can modify the foregoing embodiments and variations to incorporate various types of alternative, additional, or alternative materials. It will be understood. Accordingly, even though only a few variations of the invention are described herein, the implementation of such additional modifications and variations and equivalents thereof is defined in the following claims. It should be understood that it is within the spirit and scope of the present invention. All patent applications, patents and other publications cited herein are hereby incorporated by reference in their entirety.

Claims (23)

  A method for treating methicillin-resistant Staphylococcus aureus (MRSA) infection, wherein a subject is treated with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10 , 12,12a-Tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2- Administering a carboxylic acid amide or a pharmaceutically acceptable salt thereof.   2. The method of claim 1, wherein the pharmaceutically acceptable salt is selected from the group consisting of crystalline monohydrochloride, crystalline monomesylate, and crystalline monosulfate.   The method of claim 2, wherein the salt is a monohydrochloride salt.   3. A method according to claim 2, wherein the salt is a monomesylate.   The method of claim 2, wherein the salt is a monosulfate.   2. The method of claim 1, wherein the MRSA is selected from community-acquired MRSA (MRSA-CA) and nosocomial MRSA (MRSA-HA).   A method for treating peptic ulcer, wherein a subject is treated with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a-tetrahydroxy-7- [ (Methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or a pharmaceutically acceptable salt thereof Administering a salt.   8. The method of claim 7, wherein the pharmaceutically acceptable salt is selected from the group consisting of crystalline monohydrochloride, crystalline monomesylate, and crystalline monosulfate.   9. The method of claim 8, wherein the salt is a monohydrochloride salt.   9. The method of claim 8, wherein the salt is a monomesylate.   9. The method of claim 8, wherein the salt is monosulfate.   8. The method of claim 7, wherein the method further comprises administering at least one additional active ingredient to the subject.   13. The method of claim 12, wherein the at least one additional active ingredient is selected from a proton pump inhibitor and bismuth.   A method for treating Helicobacter pylori infection, wherein a subject is treated with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a- Tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or its Administering a pharmaceutically acceptable salt.   15. The method of claim 14, wherein the pharmaceutically acceptable salt is selected from the group consisting of crystalline monohydrochloride, crystalline monomesylate, and crystalline monosulfate.   16. The method of claim 15, wherein the salt is a monohydrochloride salt.   16. The method of claim 15, wherein the salt is a monomesylate.   16. The method of claim 15, wherein the salt is monosulfate.   A method for treating Chlamydia trachomatis infection, wherein a subject is treated with a therapeutically effective amount of (4S, 4aS, 5aR, 12aS) -4-dimethylamino-3,10,12,12a- Tetrahydroxy-7-[(methoxy (methyl) amino) -methyl] -1,11-dioxo-1,4,4a, 5,5a, 6,11,12a-octahydro-naphthacene-2-carboxylic acid amide or its Administering a pharmaceutically acceptable salt.   20. The method of claim 19, wherein the pharmaceutically acceptable salt is selected from the group consisting of crystalline monohydrochloride, crystalline monomesylate, and crystalline monosulfate.   21. The method of claim 20, wherein the salt is a monohydrochloride salt.   21. The method of claim 20, wherein the salt is a monomesylate.   21. The method of claim 20, wherein the salt is monosulfate.

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